Figure 1: Plate #1 contains competent E. coli BL21 (DE3) bacteria with pGEM-gbr22 DNA plasmid on SOC+Amp media to serve as the experimental group. Plate #2 contains competent E. coli BL21 (DE3) bacteria without the pGEM-gbr22 DNA plasmid on SOC+Amp media to serve as negative control. Plate #3 is the "fun plate" with bacterial samples from my belly button on LB+Amp media. All plates were placed in an incubator at 37 degrees Celsius for approximately 12 hours. Small white dots are indicative of colonies of bacterial growth.
Figure 2: Both 125mL Erlenmeyer flasks contain competent E. Coli BL21 (DE3) bacteria with pGEM-gbr22 DNA plasmid in LB media. These bacterial cultures were grown from bacterial colonies taken from plates of competent E. Coli BL21(DE3) bacteria with pGEM-gbr22 DNA plasmid on SOC+Amp media. The cultures were placed in a shaking incubator at 37 degrees Celsius and 250-300 rpm for approximately 16 hours. The purple color of the cultures indicates that the bacteria are producing the purple proteins coded for in the plasmid DNA, and thus they have been transformed.
Figure 3: Both 50mL conical tubes contain wet pellets of competent E. Coli BL21 (DE3) bacteria with pGEM-gbr22 DNA plasmid. The samples had been centrifuged for 10 minutes at 5000 rpm at 4 degrees Celsius. The remaining liquid was then decanted.
Figure 4: Elutions #1 and #1 of pgbr22 released from Ni-NTA chromatography resin with 1xPBS and 250mM imidazole elution buffer.
Figure 5: Dry electrophoresis gel with cellophane covering. From left to right: protein ladder, cell lysate, soluble fraction, flow through, wash, Elution 1, Elution 2, protein ladder. Discussion:
Introduction:
Materials & Methods:
Results:
Figure 1: Plate #1 contains competent E. coli BL21 (DE3) bacteria with pGEM-gbr22 DNA plasmid on SOC+Amp media to serve as the experimental group. Plate #2 contains competent E. coli BL21 (DE3) bacteria without the pGEM-gbr22 DNA plasmid on SOC+Amp media to serve as negative control. Plate #3 is the "fun plate" with bacterial samples from my belly button on LB+Amp media. All plates were placed in an incubator at 37 degrees Celsius for approximately 12 hours. Small white dots are indicative of colonies of bacterial growth.
Figure 2: Both 125mL Erlenmeyer flasks contain competent E. Coli BL21 (DE3) bacteria with pGEM-gbr22 DNA plasmid in LB media. These bacterial cultures were grown from bacterial colonies taken from plates of competent E. Coli BL21(DE3) bacteria with pGEM-gbr22 DNA plasmid on SOC+Amp media. The cultures were placed in a shaking incubator at 37 degrees Celsius and 250-300 rpm for approximately 16 hours. The purple color of the cultures indicates that the bacteria are producing the purple proteins coded for in the plasmid DNA, and thus they have been transformed.
Figure 3: Both 50mL conical tubes contain wet pellets of competent E. Coli BL21 (DE3) bacteria with pGEM-gbr22 DNA plasmid. The samples had been centrifuged for 10 minutes at 5000 rpm at 4 degrees Celsius. The remaining liquid was then decanted.
Figure 4: Elutions #1 and #1 of pgbr22 released from Ni-NTA chromatography resin with 1xPBS and 250mM imidazole elution buffer.
Figure 5: Dry electrophoresis gel with cellophane covering. From left to right: protein ladder, cell lysate, soluble fraction, flow through, wash, Elution 1, Elution 2, protein ladder.
Discussion:
Conclusions:
References: